DNA curtains provide high-throughput tools for single molecule imaging
Recently there has been a dramatic increase in the use of technologies that enable detailed interrogation of individual biological macromolecules in near-native conditions. Single molecule visualization techniques can reveal details of reaction mechanisms and macromolecular dynamics inaccessible to traditional biochemical assays. However, these techniques are often limited by the inherent challenges of collecting statistically relevant information from experiments designed to look at single events. New approaches that increase throughput capacity of single molecule methods have high potential for making these techniques readily applicable to a variety of biological questions, such as different types of DNA transactions.
DNA molecules organized into curtains along nanofabricated chromium barriers for molecule imaging
This technology is based on the discovery that nucleic acid molecules can be disposed on a substrate and positionally aligned to allow analysis of individual nucleic acid molecules. This technology provides an array that can organize DNA molecules into curtains along the leading edges of nanofabricated chromium barriers, which are located at strategic positions on a fused silica slide otherwise coated with a supported lipid bilayer. The individual molecules that make up the DNA curtains are visualized by total internal reflection fluorescence (TIRFM) allowing simultaneous imaging of thousands of perfectly aligned molecules.
Lead Inventor:
Eric C. Greene, Ph.D.
Applications:
New tools to facilitate massively parallel data collection for single molecule analysis of protein-nucleic acid interaction, protein folding and unfolding, DNA dynamics
New tools to generate maps of binding sites for DNA-binding protein that can be tagged with fluorescent labels
Offers platform for developing automated image analysis
Advantages:
A robust and powerful platform portending massively parallel data acquisition of individual protein-nucleic acid interactions in real time
Perfect alignment of DNA molecules within the curtains facilitates data evaluation